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Background of mGluR4 / GRM4 antibody

Metabotropic glutamate receptors (mGluRs) belong to the super family of G-protein coupled receptors (seven transmembrane proteins). mGluRs are further divided into subfamilies: group I mGluRs (mGluR1 and mGluR5) which couple to Gq, thereby activating phospholipase C (PLC). Group II which include mGluR2 and mGluR3 couple to Gi, therefore inhibit the formation of adenylate cyclase. mGluR4, 6, 7, 8 which belong to group III also inhibit adenylate cyclase formation by coupling to Gi. The C-terminus of these receptors has important functions in modulating their activity. This region is important for G-protein coupling, post-translatiol modifications like phosphorylation as well as protein-protein interactions. The C-termil region is also subject to altertive splicing. mGluR4 is predomintly expressed presyptically in neurons and in the cerebellum. A splice variant of the protein is expressed in taste buds. This variant lacks a large portion of the N-terminus and is normally referred to taste mGluR4. It is responsible (along with taste mGluR1) for mediating the taste of monosodium glutamate (or umi). mGluR4 knock out mice display impaired cerebellar syptic plasticity as well as some learning disabilities. In addition, this receptor has emerged as a target for the treatment of Parkinsonâ??s disease.

Immunohistochemical analysis of mGLUR4 staining in human brain formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of mGLUR4 staining in HeLa cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 C in a humidified chamber. Cells were washed with PBST and incubated with a DyLight 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).